Time-sensitive Collection and Analysis of a Marine-Derived Toxic Event Harmful algal blooms (HABs) impact many coastal states over large geographic areas and threaten environmental and socio-economical resources as well as human health and well-being. This project aims to use pre-bloom, bloom and post-bloom filtered samples from the current, September 2013, southern California bloom of Lingulodinium polyedrum to create a reference genome and transcriptome database, and assess toxin production and effect in human cell assays. The unpredictable timing of these blooms makes the current bloom event an opportunity to collect concentrated samples to evaluate potential harm to human health during these episodic events. The samples will be used for DNA and RNA sequencing and for secondary metabolite extractions, the information created will be placed in a reference library as the basis for further examination of archived samples. As part of ongoing collections, physical parameters will be measured and integrated with the genomic, transcriptomic and secondary metabolomic reference library to further aid in interpreting the data. Aim 1 describes the collections made and collections currently being performed. Aim 2 addresses the extraction, sequencing, assembly and annotation of DNA and RNA from these samples. This information will be coupled with a chemistry effort (Aim 3a) where samples will be investigated for their production of secondary metabolites (potential toxins, including sub-lethal) using mass spectrometry and NMR methods. The isolated secondary metabolites will be evaluated in four phenotypic in vitro assays used as a proxy for their potential cytotoxicity, neuro- and immuno-modulation even at sub-lethal levels (Aim 3b). In total, the work will provide fundamental knowledge by creating a reference library that will aid in further examination of archived samples collected since 2008, containing both bloom and non-bloom samples. This collected information is essential for evaluation of mitigation strategies and to provide early detection of toxic events in order to limit the impacts on human health, and will serve as the basis of an R01 grant application to be submitted next year.